Team:Ulink-SIP/lab/experimentalstep
Experimental Steps
RcsA and RcsB protein are added to overexpressed the gene manA, manB, manC, gmd, wcaG. manA gene produced protein turn fructose-6-P into Mannose-6-P. manB gene produce protein turn Mannose-6-P into Mannose-1-P. manC gene produce protein turns Mannose-1-P into GDP-Mannose. nudK gene is inhibited to prevent GDP-Mannose to be turned back into Mannose-6-P. nudD gene is inhibited in order to avoid GDP-Mannose to be broken down into GDP and Mannose. gmd gene is to turn GDP-Mannose into GDP-4-keto-6-deoxymannose. wcaG gene is used to react GDP-4-keto-6-deoxymannose and NADPH into GDP-L-fucose and NADP+. wcaj gene is inhibited to prevent it been turned into colanic acid. LacY gene is introduced into the bacteria to make channel proteins for lactose to come into the bacteria. Once the lactose come inside the bacteria, the lacZ gene needs to be inhibited to prevent the breakdown of lactose. The last step, futC gene, is overexpressed to combine lactose and GDP-L-fucose into 2’-Fucosyllactose.